微小RNA-98-5p靶向调控核糖核苷酸还原酶小亚基M2调控宫颈癌细胞顺铂的耐药性  

MicroRNA-98-5p targeting ribonucleotide reductase small subunit M2 regulating cisplatin resistance in cervical cancer cells

作　　者：张霞 蒋志明 程晓燕 项艳 赵晓 黄玉萍 于皓 ZHANG Xia;JIANG Zhi-ming;CHENG Xiao-yan;XIANG Yan;ZHAO Xiao;HUANG Yu-ping;YU Hao(Department of Gynecology,Tianxiang Oriental Hospital,Zhejiang Yiwu 322099,China;Department of Gynecology,Lishui maternal and child health hospital,Zhejiang Lishui 323000,China)

机构地区：[1]浙江义乌天祥东方医院妇科,浙江义乌322099 [2]丽水市妇幼保健院妇科,浙江丽水323000

出　　处：《解剖学报》2021年第3期432-438,共7页Acta Anatomica Sinica

基　　金：丽水市科技计划项目(2019GYX22)。

摘　　要：目的探讨微小RNA(miR)-98-5p对顺铂(DDP)耐药宫颈癌细胞顺铂敏感性的调控及其机制。方法用脂质体法将DDP+miR-NC组(转染miR-NC)、DDP+miR-98-5p组(转染miR-98-5p mimics)、DDP+si-NC组(转染si-NC)、DDP+si-核糖核苷酸还原酶小亚基M2(RRM2)组(转染si-RRM2)、DDP+miR-98-5p+pc DNA组(共转染miR-98-5p mimics和pc DNA)、DDP+miR-98-5p+pc DNA-RRM2组(共转染miR-98-5p mimics和pc DNA-RRM2)转染至He La/DDP组细胞;Real-time PCR、Western blotting、CCK-8、迁移实验(Transwell)和双荧光素酶报告基因检测细胞中miR-98-5p、RRM2、细胞周期蛋白D1(cyclin D1)、P21、基因金属蛋白酶(MMP)-2、MMP-9的表达、细胞的抑制率、半数抑制浓度(IC50)、迁移和侵袭及荧光活性。结果与He La组细胞相比,He La/DDP组细胞中miR-98-5p表达显著降低,RRM2表达显著升高,IC50值显著升高(P<0.05);过表达miR-98-5p或抑制RRM2可明显抑制He La/DDP细胞的增殖、迁移和侵袭,下调cyclin D1、MMP-2、MMP-9蛋白,上调P21蛋白;miR-98-5p明显抑制野生型RRM2细胞的荧光活性,并且过表达RRM2反转了miR-98-5p对He La/DDP细胞增殖、迁移侵袭的抑制作用。结论MiR-98-5p可抑制顺铂耐药宫颈癌细胞的增殖、迁移和侵袭,增强对顺铂的敏感性,其机制与靶向RRM2相关,将可为顺铂耐药宫颈癌细胞的治疗提供方向。Objective To investigate the regulation and mechanism of microRNA(miR)-98-5p on cisplatin sensitivity in cisplatin-resistant cervical cancer cells.Methods The cisplatin(DDP) + miR-NC group(transfected miRNC),DDP + miR-98-5p group(transfected miR-98-5p mimics),DDP + si-NC group(transfected si-NC),DDP + siribonucleotide reductase subunit M2(RRM2) group(transfected si-RRM2),DDP + miR-98-5p + pc DNA group(cotransfected miR-98-5p mimics and pc DNA),DDP + miR-98-5p + pc DNA-RRM2 group(co-transfected miR-98-5p mimics and pc DNA-RRM2) were transfected into He La/DDP cells by liposome method.Real-tim PCR,Western blotting,CCK-8,Transwell chamber and dual luciferase reports gene detection assay were used to detect the expression of miR-98-5p,RRM2,cyclin D1, P21, matrix metalloproteinase(MMP)-2 and MMP-9 in cells, inhibition rate, half inhibitory concentration(IC50),migration and invasion and fluorescence activity.Results Compared with the He La group,the expression of miR-98-5p was significantly decreased in He La/DDP group,the expression of RRM2 was significantly increased,the IC50 value was significantly increased(P < 0.05).Overexpression miR-98-5p or inhibition RRM2 could significantly inhibit the proliferation,migration and invasion of He La/DDP cells,down-regulated proteins expression of cyclin D1,MMP-2 and MMP-9,and up-regulated proteins expression of P21;miR-98-5p inhibited the fluorescence activity of wild-type RRM2 cells,and overexpress RRM2 could reverse the inhibitory effect of miR-98-5p on proliferation,migration and invasion of He La/DDP cells.Conclusion miR-98-5p can inhibit the proliferation,migration and invasion of cisplatinresistant cervical cancer cells and enhance the sensitivity to cisplatin.The mechanism is related to the targeting of RRM2,which will provide directions for the treatment of cisplatin-resistant cervical cancer cells.

关 键 词：宫颈癌 微小RNA-98-5p 核糖核苷酸还原酶小亚基M2 顺铂 免疫印迹法 人 

分 类 号：R737.33[医药卫生—肿瘤]